Methods of administering platelet derivative compositions to reduce bleeding in subjects refractory to platelet transfusion

Abstract

Provided herein are compositions, and methods for administering a platelet derivative composition for reducing bleeding in a subject who is refractory to platelet transfusion. Methods herein can further include administering a platelet derivative composition to a subject having low platelet counts (e.g., having thrombocytopenia) and/or to maintain the number of platelets over a threshold amount, or to increase the number of platelets in the subject. Methods herein can include administering a therapeutic agent before and optionally after administering the platelet derivative composition to the subject. Methods herein can reduce or eliminate any dosing and/or scheduling adjustments of a therapeutic regimen due to a low platelet count.

What is claimed is: 
     
         1 . A method of decreasing bleeding of a subject, comprising:
 rehydrating a platelet derivative composition in the form of a powder to form a rehydrated platelet derivative composition comprising rehydrated platelet derivatives, and   administering a dose of the rehydrated platelet derivative composition to the subject, wherein the dose comprises a first dose of the rehydrated platelet derivatives,   wherein at least 70% of the rehydrated platelet derivatives in the rehydrated platelet derivative composition are CD41 positive platelet derivatives when measured using flow cytometry, and less than 5% of the CD41 positive platelet derivatives are microparticles,   wherein the rehydrated platelet derivatives in the rehydrated platelet derivative composition have a compromised membrane,   wherein at least 70% of the rehydrated platelet derivatives in the rehydrated platelet derivative composition are CD42 positive platelet derivatives, when measured using flow cytometry,   wherein at least 70% of the rehydrated platelet derivatives in the rehydrated platelet derivative composition are CD62 positive platelet derivatives, when measured using flow cytometry,   wherein the rehydrated platelet derivatives are capable of generating thrombin in an in vitro thrombin formation assay,   wherein the rehydrated platelet derivatives show an inability to increase expression of a platelet activation marker in the presence of an agonist as compared to the expression of the platelet activation marker in the absence of an agonist, and wherein the platelet activation marker is Annexin V, CD62, or a combination thereof,   wherein the subject is refractory to platelet transfusion,   wherein the first dose of the rehydrated platelet derivatives is an amount of between 5.1×10 8  and 9.9×10 8 /kg, or 1.0×10 9  and 1.0×10 10 /kg of the subject, and   wherein the administering leads to the subject having decreased bleeding without a bleeding score of WHO Grade 2A or greater at 12 hours after the administering the first dose of the platelet derivatives.   
     
     
         2 . The method of  claim 1 , wherein the subject has thrombocytopenia at the time of the administering the dose. 
     
     
         3 . The method of  claim 2 , wherein the subject at the time of the administering the first dose of the rehydrated platelet derivatives has a bleeding score of WHO grade 2 but not solely the result of cutaneous bleeding. 
     
     
         4 . The method of  claim 1 , wherein the first dose of the rehydrated platelet derivatives is an amount of between 1.0×10 9  and 1.0×10 10 /kg of the subject. 
     
     
         5 . The method of  claim 1 , wherein the administering the dose, comprises administering more than 1 dose to a maximum of 10 doses of the rehydrated platelet derivative composition, wherein each dose of the maximum of 10 doses is an amount of rehydrated platelet derivatives between 5.1×10 8  and 9.9×10 8 /kg, or 1.0×10 9  and 1.0×10 10 /kg of the subject. 
     
     
         6 . The method of  claim 3 , wherein the subject has decreased bleeding without a bleeding score of WHO Grade 2A or greater at 24 hours after the administering the first dose of the platelet derivatives. 
     
     
         7 . A method of decreasing bleeding of a subject, comprising:
 rehydrating a platelet derivative composition in the form of a powder to form a rehydrated platelet derivative composition comprising rehydrated platelet derivatives, and   administering a dose of the rehydrated platelet derivative composition to the subject, wherein the dose comprises a first dose of the rehydrated platelet derivatives,   wherein at least 70% of the rehydrated platelet derivatives in the rehydrated platelet derivative composition are CD41 positive platelet derivatives when measured using flow cytometry, and less than 5% of the CD41 positive platelet derivatives are microparticles,   wherein the rehydrated platelet derivatives in the rehydrated platelet derivative composition have a compromised membrane,   wherein at least 70% of the rehydrated platelet derivatives in the rehydrated platelet derivative composition are CD42 positive platelet derivatives, when measured using flow cytometry,   wherein at least 70% of the rehydrated platelet derivatives in the rehydrated platelet derivative composition are CD62 positive platelet derivatives, when measured using flow cytometry,   wherein the rehydrated platelet derivatives are capable of generating thrombin in an in vitro thrombin formation assay,   wherein the rehydrated platelet derivatives show an inability to increase expression of a platelet activation marker in the presence of an agonist as compared to the expression of the platelet activation marker in the absence of an agonist, and wherein the platelet activation marker is Annexin V, CD62, or a combination thereof,   wherein the subject is refractory to platelet transfusion,   wherein the first dose of the rehydrated platelet derivatives is an amount of between 5×10 8  particles/kg and 1.0×10 12  particles/kg of the subject, and   wherein the administering leads to the subject having decreased bleeding without a bleeding score of WHO Grade 2A or greater at 12 hours after the administering the first dose of the platelet derivatives.   
     
     
         8 . The method of  claim 7 , wherein the subject has thrombocytopenia at the time of the administering the dose. 
     
     
         9 . The method of  claim 7 , wherein the subject has severe thrombocytopenia at the time of the administering the dose. 
     
     
         10 . The method of  claim 7 , wherein the first dose of the rehydrated platelet derivatives is an amount of between 5.1×10 8  and 9.9×10 8 /kg of the subject. 
     
     
         11 . The method of  claim 7 , wherein the first dose of the rehydrated platelet derivatives is an amount of between 1.0×10 9  and 1.0×10 12 /kg of the subject. 
     
     
         12 . The method of  claim 7 , wherein the first dose of the rehydrated platelet derivatives is an amount of between 1.0×10 9  and 1.0×10 10 /kg of the subject. 
     
     
         13 . The method of  claim 7 , wherein the administering the dose, comprises administering between 2 and 10 doses of the rehydrated platelet derivative composition, wherein each dose that is administered between 2 and 10 times is an amount of rehydrated platelet derivatives between 5×10 8  particles/kg and 1.0×10 12  particles/kg of the subject. 
     
     
         14 . The method of  claim 7 , wherein the subject at the time of the administering the first dose of the rehydrated platelet derivatives has a bleeding score of WHO grade 2 but not solely the result of cutaneous bleeding. 
     
     
         15 . The method of  claim 14 , wherein the subject has decreased bleeding without a bleeding score of WHO Grade 2A or greater at 24 hours after the administering the first dose of the rehydrated platelet derivatives. 
     
     
         16 . The method of  claim 7 , wherein the subject has anti-human leukocyte antigen (HLA) antibodies and/or anti-human platelet antigen (HPA) antibodies in their blood at the time of the administering the first dose of the rehydrated platelet derivatives. 
     
     
         17 . The method of  claim 7 , wherein the subject has cross-reactive antibodies against the rehydrated platelet derivatives. 
     
     
         18 . The method of  claim 7 , wherein the administering the dose, comprises administering between 2 and 8 times the dose of the rehydrated platelet derivative composition, wherein each dose that is administered between 2 and 8 times is an amount of rehydrated platelet derivatives between 5×10 8  particles/kg and 1.0×10 12  particles/kg of the subject. 
     
     
         19 . The method of  claim 7 , wherein the subject experiences no adverse events attributable to the administering the dose of the rehydrated platelet derivative composition when the decreased bleeding is detected. 
     
     
         20 . The method of  claim 14 , wherein within 3 days after the administering the first dose of the rehydrated platelet derivatives, the bleeding of the subject is decreased to a WHO bleeding score of 0. 
     
     
         21 . The method of  claim 7 , wherein the administering the dose, comprises administering between 2 and 6 times the dose of the rehydrated platelet derivative composition, wherein each dose that is administered between 2 and 6 times is an amount of rehydrated platelet derivatives between 5×10 8  particles/kg and 1.0×10 12  particles/kg of the subject. 
     
     
         22 . The method of  claim 7 , wherein at the time of the administering the first dose of the rehydrated platelet derivatives, the subject has a WHO bleeding score of Grade 2 or Grade 3. 
     
     
         23 . The method of any one of the other claims herein, wherein the administering the dose of the rehydrated platelet derivative composition restores normal hemostasis in the subject. 
     
     
         24 . The method of  claim 7 , wherein the subject has Glanzmann thrombasthenia. 
     
     
         25 . The method of  claim 7 , wherein the subject is not undergoing another treatment for thrombocytopenia at the time between the administering the first dose of the rehydrated platelet derivatives and a determination that the bleeding is reduced. 
     
     
         26 . The method of  claim 7 , wherein at the time of administering the first dose of the rehydrated platelet derivatives, the subject does not have another thrombotic or ischemic condition. 
     
     
         27 . The method of  claim 7 , wherein at the time of administering the first dose of the rehydrated platelet derivatives, the subject is not taking any pro or anti-thrombotic medications. 
     
     
         28 . The method of  claim 14 , wherein after the administering the dose of the rehydrated platelet derivative composition, the subject has no sites that are actively bleeding with a bleeding score WHO Grade 2A or greater. 
     
     
         29 . The method of  claim 7 , wherein the administering the dose of the rehydrated platelet derivative composition leads to cessation of bleeding at the primary bleeding site. 
     
     
         30 . The method of  claim 29 , wherein the administering the dose of the rehydrated platelet derivative composition leads to cessation of bleeding at all bleeding sites other than the primary bleeding site. 
     
     
         31 . The method of  claim 7 , wherein the administering the dose of the rehydrated platelet derivative composition is performed until the bleeding stops at a primary bleeding site. 
     
     
         32 . The method of  claim 31 , wherein the administering the dose of the rehydrated platelet derivative composition is performed until there is cessation of bleeding at a bleeding site other than the primary bleeding site. 
     
     
         33 . The method of  claim 31 , wherein the administering the dose of the rehydrated platelet derivative composition is performed until there is cessation of bleeding at all bleeding sites in the subject. 
     
     
         34 . The method of any one of the other claims herein, wherein the rehydrated platelet derivative composition comprises less than 3% microparticles. 
     
     
         35 . The method of any one of the other claims herein, wherein at least 75% of the rehydrated platelet derivatives are CD41-positive, at least 75% of the rehydrated platelet derivatives are CD42-positive, at least 75% of the rehydrated platelet derivatives are CD62-positive. 
     
     
         36 . The method of any one of the other claims herein, wherein the rehydrated platelet derivatives have one or both of:
 thrombospondin (TSP) on their surface at a level that is at least 50% higher than on the surface of resting platelets; and   von Willebrand factor (vWF) on their surface at a level that is at least 50% higher than on the surface of resting platelets.   
     
     
         37 . The method of any one of the other claims herein, wherein the rehydrated platelet derivative composition comprises microparticles having a diameter less than 0.5 μm and present in an amount in the range of 0.1 to 4.9%. 
     
     
         38 . The method of any one of the other claims herein, wherein the administering the dose is performed by infusing the rehydrated platelet derivative composition to the subject. 
     
     
         39 . The method of any one of the other claims herein, wherein the administering the dose leads to a cessation or a decrease in bleeding at a primary bleeding site at 12 hours, or 24 hours after the administering the first dose of the rehydrated platelet derivatives. 
     
     
         40 . The method of any one of the other claims herein, wherein the administering the dose leads to a cessation or a decrease in bleeding at bleeding sites other than a primary bleeding site at 12 hours, or 24 hours after the administering the first dose of the rehydrated platelet derivatives. 
     
     
         41 . The method of  claim 7 , wherein the administering the dose leads to cessation of bleeding at all bleeding sites within 12, or 24 hours after the administering the first dose of the rehydrated platelet derivatives. 
     
     
         42 . The method of  claim 7 , wherein the subject experiences no adverse events, or no grade 2 or higher adverse events, attributable to the administering the dose of the rehydrated platelet derivative composition when the decreased bleeding is detected. 
     
     
         43 . The method of  claim 7 , wherein the subject experiences no grade 3 or grade 4 adverse events attributable to the administering the dose of the rehydrated platelet derivative composition, when the decreased bleeding is detected. 
     
     
         44 . The method of  claim 7 , wherein the subject experiences no adverse events or no grade 2 or higher adverse events, attributable to the administering the dose of the rehydrated platelet derivative composition, within 30 days of the first dose. 
     
     
         45 . The method of  claim 7 , wherein the subject experiences no grade 2 or higher adverse events, attributable to the administering the dose of the rehydrated platelet derivative composition, within 24 hours of the first dose. 
     
     
         46 . The method of  claim 7 , wherein the subject experiences no adverse events, or no grade 2 or higher adverse events, attributable to the administering the dose of the rehydrated platelet derivative composition, within 72 hours of the first dose. 
     
     
         47 . The method of  claim 7 , wherein the subject experiences no adverse events, or no grade 2 or higher adverse events, attributable to the administering the dose of the rehydrated platelet derivative composition, within 12 hours of the first dose. 
     
     
         48 . The method of any one of the other claims herein, wherein the rehydrated platelet derivatives when at a concentration of about 255×10 3  platelet derivatives/μL are capable of producing occlusion by attaining a pressure of 80 kPa in less than 14 minutes in platelet-reduced citrated whole blood in an in vitro total thrombus-formation analysis system (T-TAS). 
     
     
         49 . The method of  claim 7 , wherein the rehydrated platelet derivatives are capable of yielding+/−25% of one or more occlusion start time, occlusion time, or area under the curve (AUC) in an in vitro T-TAS assay in the presence versus absence of anti-HLA and/or anti-HPA antibodies. 
     
     
         50 . The method of  claim 7 , wherein the rehydrated platelet derivatives are capable of yielding+/−25% of one or more occlusion start time, occlusion time, or area under the curve (AUC) in an in vitro T-TAS assay in the presence versus absence of anti-HPA antibodies. 
     
     
         51 . The method of  claim 7 , wherein the rehydrated platelet derivatives restore thrombin levels identical 4,800 particles/microliter. 
     
     
         52 . The method of  claim 7 , wherein the rehydrated platelet derivatives are capable of yielding comparable in vitro thrombin generation activity in the presence versus absence of cross-reactive antibodies against the platelet derivatives. 
     
     
         53 . The method of  claim 7 , wherein the rehydrated platelet derivatives are capable of yielding comparable hemostatic activity in an in vitro T-TAS assay in the presence versus absence of cross-reactive antibodies against the platelet derivatives. 
     
     
         54 . The method of  claim 7 , wherein the rehydrated platelet derivatives are capable of yielding comparable in vitro thrombin generation activity in the presence versus absence of anti-HLA and/or anti-HPA antibodies. 
     
     
         55 . The method of  claim 7 , wherein the rehydrated platelet derivatives are capable of yielding comparable hemostatic activity in an in vitro T-TAS assay in the presence versus absence of anti-HLA and/or anti-HPA antibodies. 
     
     
         56 . The method of  claim 7 , wherein the administering the dose of the rehydrated platelet derivative composition leads to an improvement in thrombin generation in the subject as compared to the subject before the administering and/or an improvement in clot formation in the subject as compared to the subject before the administering. 
     
     
         57 . The method of  claim 7 , wherein the rehydrated platelet derivatives have 12% to 64%, 20% to 64%, 20% to 60%, 20% to 50%, or 25% to 50% amount of CD42 as compared to the amount of CD42 in fresh platelets or apheresis platelets.